Forough Reza, Weylie Brian, Patel Chirag, Ambrus Sandy, Singh Ugra S, Zhu James
Department of Medical Physiology and Cardiovascular Research Institute, College of Medicine, The Texas A&M University System Health Science Center, College Station, Texas 77843, USA.
J Cell Biochem. 2005 Jan 1;94(1):109-16. doi: 10.1002/jcb.20274.
Transfection of chicken chorioallantoic membranes (CAMs) with a chimeric secreted version of fibroblast growth factor-1 (sp-FGF-1) gene construct leads to a significant increase in vascularization. Though FGF-stimulated angiogenesis has been extensively studied, the molecular mechanisms regulating FGF-1-induced angiogenesis are poorly understood in vivo. This study was designed to investigate the role of the AKT (PKB) kinase signaling pathway in mediating sp-FGF-1-induced angiogenesis in the chicken CAM. The involvement of the AKT pathway was demonstrated by up-regulation of AKT1 mRNA expression in sp-FGF-1 compared to vector alone control transfected CAMs as demonstrated by real-time RT-PCR. Western analysis using an antibody specific to the activated AKT (phosphorylated AKT), demonstrated an increase in AKT activity in sp-FGF-1 compared to vector control transfected CAMs. More importantly, the AKT inhibitor ML-9 significantly reduced sp-FGF-1-induced angiogenesis in CAMs. These results indicate that AKT signaling plays a role in FGF-1-stimulated angiogenesis in vivo and the AKT pathway may serve as a therapeutic target for angiogenesis-associated diseases.
用成纤维细胞生长因子-1（sp-FGF-1）基因构建体的嵌合分泌型转染鸡绒毛尿囊膜（CAMs）会导致血管生成显著增加。尽管FGF刺激的血管生成已得到广泛研究，但在体内调节FGF-1诱导血管生成的分子机制仍知之甚少。本研究旨在探讨AKT（PKB）激酶信号通路在介导sp-FGF-1诱导鸡CAM血管生成中的作用。通过实时RT-PCR证明，与单独载体对照转染的CAMs相比，sp-FGF-1中AKT1 mRNA表达上调，从而证明了AKT途径的参与。使用针对活化AKT（磷酸化AKT）的特异性抗体进行的蛋白质印迹分析表明，与载体对照转染的CAMs相比，sp-FGF-1中AKT活性增加。更重要的是，AKT抑制剂ML-9显著降低了sp-FGF-1诱导的CAM血管生成。这些结果表明，AKT信号在体内FGF-1刺激的血管生成中起作用，并且AKT途径可能作为血管生成相关疾病的治疗靶点。
